Buffer solution

A buffer solution (more precisely, pH buffer or hydrogen ion buffer) is an aqueous solution consisting of a
mixture of a weak acid and its conjugate base, or vice versa. Its pH changes very little when a small amount of
strong acid or base is added to it. Buffer solutions are used as a means of keeping pH at a nearly constant value
in a wide variety of chemical applications. In nature, there are many systems that use buffering for pH
regulation. For example, the bicarbonate buffering system is used to regulate the pH of blood.

Contents
Principles of buffering
Buffer capacity
Applications
Simple buffering agents
"Universal" buffer mixtures
Common buffer compounds used in biology
Calculating buffer pH
Monoprotic acids
Polyprotic acids
See also
References
External links

Principles of buffering
Buffer solutions achieve their resistance to pH change because of
the presence of an equilibrium between the weak acid HA and its
conjugate base A−:

HA ⇌ H+ + A−

When some strong acid is added to an equilibrium mixture of the

weak acid and its conjugate base, hydrogen ions (H+) are added,
and the equilibrium is shifted to the left, in accordance with Le
Châtelier's principle. Because of this, the hydrogen ion
concentration increases by less than the amount expected for the
quantity of strong acid added. Similarly, if strong alkali is added
to the mixture, the hydrogen ion concentration decreases by less Simulated titration of an acidified solution
than the amount expected for the quantity of alkali added. The of a weak acid (pKa = 4.7) with alkali
effect is illustrated by the simulated titration of a weak acid with
pKa = 4.7. The relative concentration of undissociated acid is
shown in blue, and of its conjugate base in red. The pH changes relatively slowly in the buffer region,
pH = pKa ± 1, centered at pH = 4.7, where [HA] = [A−]. The hydrogen ion concentration decreases by less
than the amount expected because most of the added hydroxide ion is consumed in the reaction

OH− + HA → H2O + A−

and only a little is consumed in the neutralization reaction (which is the reaction that results in an increase in
pH)

OH− + H+ → H2O.

Once the acid is more than 95% deprotonated, the pH rises rapidly because most of the added alkali is
consumed in the neutralization reaction.

Buffer capacity

Buffer capacity is a quantitative measure of the resistance to change of pH of a solution containing a buffering
agent with respect to a change of acid or alkali concentration. It can be defined as follows:[1][2]

where is an infinitesimal amount of added base, or

where is an infinitesimal amount of added acid. pH is defined as −log10 [H+], and d(pH) is an
infinitesimal change in pH.

With either definition the buffer capacity for a weak acid HA with dissociation constant Ka can be expressed
as[3][4][2]

where [H+] is the concentration of hydrogen ions, and is the total concentration of added acid. Kw is the
equilibrium constant for self-ionization of water, equal to 1.0 × 10−14 . Note that in solution H+ exists as the
hydronium ion H3 O+, and further aquation of the hydronium ion has negligible effect on the dissociation
equilibrium, except at very high acid concentration.

This equation shows that there are three regions of raised buffer capacity (see figure).

In the central region of the curve (coloured green on the plot), the second term is dominant, and

Buffer capacity rises to a local maximum at pH = pKa. The height of this peak depends on the
value of pKa. Buffer capacity is negligible when the concentration [HA] of buffering agent is
 very small and increases with increasing
concentration of the buffering agent.[2] Some authors
show only this region in graphs of buffer capacity.[1]
Buffer capacity falls to 33% of the maximum value at
pH = pKa ± 1, to 10% at pH = pKa ± 1.5 and to 1% at
pH = pKa ± 2. For this reason the most useful range
is approximately pKa ± 1. When choosing a buffer for
use at a specific pH, it should have a pKa value as
close as possible to that pH.[1]

With strongly acidic solutions, pH less than about 2

(coloured red on the plot), the first term in the equation
dominates, and buffer capacity rises exponentially with
decreasing pH:
Buffer capacity β for a 0.1 M solution of a
weak acid with a pKa = 7

This results from the fact that the second and third
terms become negligible at very low pH. This term is independent of the presence or
absence of a buffering agent.

With strongly alkaline solutions, pH more than about 12 (coloured blue on the plot), the third
term in the equation dominates, and buffer capacity rises exponentially with increasing pH:

This results from the fact that the first and second terms become negligible at very high pH.
This term is also independent of the presence or absence of a buffering agent.

Applications
The pH of a solution containing a buffering agent can only vary within a narrow range, regardless of what else
may be present in the solution. In biological systems this is an essential condition for enzymes to function
−
correctly. For example, in human blood a mixture of carbonic acid (H2 CO3 ) and bicarbonate (HCO3 ) is
present in the plasma fraction; this constitutes the major mechanism for maintaining the pH of blood between
7.35 and 7.45. Outside this narrow range (7.40 ± 0.05 pH unit), acidosis and alkalosis metabolic conditions
rapidly develop, ultimately leading to death if the correct buffering capacity is not rapidly restored.

If the pH value of a solution rises or falls too much, the effectiveness of an enzyme decreases in a process,
known as denaturation, which is usually irreversible.[5] The majority of biological samples that are used in
research are kept in a buffer solution, often phosphate buffered saline (PBS) at pH 7.4.

In industry, buffering agents are used in fermentation processes and in setting the correct conditions for dyes
used in colouring fabrics. They are also used in chemical analysis[4] and calibration of pH meters.

Simple buffering agents

 Buffering agent pKa Useful pH range

Citric acid 3.13, 4.76, 6.40 2.1–7.4

Acetic acid 4.8 3.8–5.8
KH2PO4 7.2 6.2–8.2

CHES 9.3 8.3–10.3

Borate 9.24 8.25–10.25

For buffers in acid regions, the pH may be adjusted to a desired value by adding a strong acid such as
hydrochloric acid to the particular buffering agent. For alkaline buffers, a strong base such as sodium
hydroxide may be added. Alternatively, a buffer mixture can be made from a mixture of an acid and its
conjugate base. For example, an acetate buffer can be made from a mixture of acetic acid and sodium acetate.
Similarly, an alkaline buffer can be made from a mixture of the base and its conjugate acid.

"Universal" buffer mixtures

By combining substances with pKa values differing by only two or less and adjusting the pH, a wide range of
buffers can be obtained. Citric acid is a useful component of a buffer mixture because it has three pKa values,
separated by less than two. The buffer range can be extended by adding other buffering agents. The following
mixtures (McIlvaine's buffer solutions) have a buffer range of pH 3 to 8.[6]

0.2 M Na2HPO4 (mL) 0.1 M citric acid (mL) pH

20.55 79.45 3.0

38.55 61.45 4.0
51.50 48.50 5.0
63.15 36.85 6.0
82.35 17.65 7.0
97.25 2.75 8.0

A mixture containing citric acid, monopotassium phosphate, boric acid, and diethyl barbituric acid can be
made to cover the pH range 2.6 to 12.[7]

Other universal buffers are the Carmody buffer[8] and the Britton–Robinson buffer, developed in 1931.

Common buffer compounds used in biology

For effective range see Buffer capacity, above.

 Temp.
pKa effect Mol.
Common name (chemical name) Structure at dpH
dT weight
25 °C
(K−1)[9]

TAPS
([tris(hydroxymethyl)methylamino]propanesulfonic 8.43 −0.018 243.3
acid)

Bicine (2-(bis(2-hydroxyethyl)amino)acetic acid) 8.35 −0.018 163.2

Tris (tris(hydroxymethyl)aminomethane) or (2-

8.07* −0.028 121.14
amino-2-(hydroxymethyl)propane-1,3-diol)

Tricine (N-[tris(hydroxymethyl)methyl]glycine) 8.05 −0.021 179.2

TAPSO (3-[N-tris(hydroxymethyl)methylamino]-2-
7.635 259.3
hydroxypropanesulfonic acid)

HEPES (4-(2-hydroxyethyl)-1-
7.48 −0.014 238.3
piperazineethanesulfonic acid)

TES (2-[[1,3-dihydroxy-2-(hydroxymethyl)propan-2-
7.40 −0.020 229.20
yl]amino]ethanesulfonic acid)

MOPS (3-(N-morpholino)propanesulfonic acid) 7.20 −0.015 209.3

PIPES (piperazine-N,N′-bis(2-ethanesulfonic acid)) 6.76 −0.008 302.4

Cacodylate (dimethylarsenic acid) 6.27 138.0

 MES (2-(N-morpholino)ethanesulfonic acid) 6.15 −0.011 195.2

(*) Tris is a base, the pKa = 8.07 refers to its conjugate acid.

Calculating buffer pH

Monoprotic acids

First write down the equilibrium expression

HA ⇌ A− + H+

This shows that when the acid dissociates, equal amounts of hydrogen ion and anion are produced. The
equilibrium concentrations of these three components can be calculated in an ICE table (ICE standing for
"initial, change, equilibrium").

ICE table for a monoprotic

acid

[HA] [A−] [H+]

I C0 0 y

C −x x x
E C0 − x x x+y

The first row, labelled I, lists the initial conditions: the concentration of acid is C0 , initially undissociated, so
the concentrations of A− and H+ would be zero; y is the initial concentration of added strong acid, such as
hydrochloric acid. If strong alkali, such as sodium hydroxide, is added, then y will have a negative sign
because alkali removes hydrogen ions from the solution. The second row, labelled C for "change", specifies
the changes that occur when the acid dissociates. The acid concentration decreases by an amount −x, and the
concentrations of A− and H+ both increase by an amount +x. This follows from the equilibrium expression.
The third row, labelled E for "equilibrium", adds together the first two rows and shows the concentrations at
equilibrium.

To find x, use the formula for the equilibrium constant in terms of concentrations:

Substitute the concentrations with the values found in the last row of the ICE table:
Simplify to

With specific values for C0 , Ka and y, this equation can be solved for x. Assuming that pH = −log10 [H+], the
pH can be calculated as pH = −log10 (x + y).

Polyprotic acids

Polyprotic acids are acids that can lose more than one proton. The
constant for dissociation of the first proton may be denoted as Ka1 ,
and the constants for dissociation of successive protons as Ka2 , etc.
Citric acid is an example of a polyprotic acid H3 A, as it can lose three
protons.

Equilibrium pKa value % species formation calculated for a

H3A ⇌ H2A− + H+ pKa1 = 3.13 10-millimolar solution of citric acid

H2A− ⇌ HA2− + H+ pKa2 = 4.76

HA2− ⇌ A3− + H+ pKa3 = 6.40

When the difference between successive pKa values is less than about 3, there is overlap between the pH
range of existence of the species in equilibrium. The smaller the difference, the more the overlap. In the case of
citric acid, the overlap is extensive and solutions of citric acid are buffered over the whole range of pH 2.5 to
7.5.

Calculation of the pH with a polyprotic acid requires a speciation calculation to be performed. In the case of
citric acid, this entails the solution of the two equations of mass balance:

CA is the analytical concentration of the acid, CH is the analytical concentration of added hydrogen ions, βq are
the cumulative association constants:

Kw is the constant for self-ionization of water. There are two non-linear simultaneous equations in two
unknown quantities [A3−] and [H+]. Many computer programs are available to do this calculation. The
speciation diagram for citric acid was produced with the program HySS.[10]

See also
Henderson–Hasselbalch equation
Buffering agent
Good's buffers
Common-ion effect
Metal ion buffer
 Mineral redox buffer

References
1. Skoog, Douglas A.; West, Donald M.; Holler, F. James; Crouch, Stanley R. (2014).
Fundamentals of Analytical Chemistry (9th ed.). Brooks/Cole. p. 226. ISBN 978-0-495-55828-6.
2. Urbansky, Edward T.; Schock, Michael R. (2000). "Understanding, Deriving and Computing
Buffer Capacity". Journal of Chemical Education. 77 (12): 1640–1644.
doi:10.1021/ed077p1640 (https://doi.org/10.1021%2Fed077p1640).
3. Butler, J. N. (1998). Ionic Equilibrium: Solubility and pH calculations. Wiley. pp. 133–136.
ISBN 978-0-471-58526-8.
4. Hulanicki, A. (1987). Reactions of acids and bases in analytical chemistry. Translated by
Masson, Mary R. Horwood. ISBN 978-0-85312-330-9.
5. Scorpio, R. (2000). Fundamentals of Acids, Bases, Buffers & Their Application to Biochemical
Systems. ISBN 978-0-7872-7374-3.
6. McIlvaine, T. C. (1921). "A buffer solution for colorimetric comparaison" (http://www.jbc.org/cont
ent/49/1/183.full.pdf) (PDF). J. Biol. Chem. 49 (1): 183–186. Archived (https://web.archive.org/w
eb/20150226111238/http://www.jbc.org/content/49/1/183.full.pdf) (PDF) from the original on
2015-02-26.
7. Mendham, J.; Denny, R. C.; Barnes, J. D.; Thomas, M. (2000). "Appendix 5". Vogel's textbook of
quantitative chemical analysis (5th ed.). Harlow: Pearson Education. ISBN 978-0-582-22628-9.
8. Carmody, Walter R. (1961). "Easily prepared wide range buffer series". J. Chem. Educ. 38 (11):
559–560. Bibcode:1961JChEd..38..559C (https://ui.adsabs.harvard.edu/abs/1961JChEd..38..5
59C). doi:10.1021/ed038p559 (https://doi.org/10.1021%2Fed038p559).
9. "Buffer Reference Center" (http://www.sigmaaldrich.com/life-science/core-bioreagents/biologic
al-buffers/learning-center/buffer-reference-center.html). Sigma-Aldrich. Archived (https://web.arc
hive.org/web/20090417003507/http://www.sigmaaldrich.com/life-science/core-bioreagents/biol
ogical-buffers/learning-center/buffer-reference-center.html) from the original on 2009-04-17.
Retrieved 2009-04-17.
10. Alderighi, L.; Gans, P.; Ienco, A.; Peters, D.; Sabatini, A.; Vacca, A. (1999). "Hyperquad
simulation and speciation (HySS): a utility program for the investigation of equilibria involving
soluble and partially soluble species" (http://www.hyperquad.co.uk/hyss.htm). Coordination
Chemistry Reviews. 184 (1): 311–318. doi:10.1016/S0010-8545(98)00260-4 (https://doi.org/10.
1016%2FS0010-8545%2898%2900260-4). Archived (https://web.archive.org/web/2007070408
3413/http://www.hyperquad.co.uk/hyss.htm) from the original on 2007-07-04.

External links
"Biological buffers" (http://www.reachdevices.com/Protein/BiologicalBuffers.html). REACH Devices.

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